New Methods to Separate Empty, Partial, Full and Heavy AAV Capsids

Learn how to achieve:

  • Five times better resolution of empty/full capsids
  • Large scale removal of empty, partial and heavy AAV capsids
  • Batch-to-batch and scale-to-scale reproducibility within +/- 3%

Speaker:
Aleš Štrancar, Sartorius BIA Separations

Abstract:
Separation of empty and full AAV capsids with very similar chromatographic characteristics requires very high-resolution chromatography media and scale-to scale reproducibility within just few percent. Qualification of chromatographic columns using small molecules might be misleading when separation of big molecules or virus particles is in question. Column release criteria based on the AAV elution profile must therefore be adopted and used for all column sizes. Recently, the presence of partial and heavy AAV capsids in gene therapy products has raised concerns due to potential carryover of host cell DNA and plasmid DNA fragments. Their removal from the product is therefore crucial. To address these needs, Sartorius BIA Separations developed columns with better resolution and narrower acceptance criteria. These columns allow for batch-to-batch and scale-to-scale reproducibility within just 3% of the AAV8 empty capsid isoconductivity elution, and step-gradient elution using the same buffer strength at any scale. Two orders of magnitude removal of empty capsid should be possible at any scale.

To address the recent needs Sartorius BIA Separations developed new CIMmultus HR line (HR stands for High Reproducibility) columns with better resolution and narrower acceptance criteria. These columns allow for step gradient elution using the same buffer strength at any scale. Two order of magnitude removal of empty capsid should be possible at any scale.

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